cirrhosis (n = 5) were stained to demonstrate OPN and analyzed using computer-assisted morphometry. Expression of OPN was lowest in patients with NAFL and highest in patients with NASH-related cirrhosis (Fig. 6A,B). To further validate the association between NAFLD-related fibrosis stage and OPN expression, total liver RNA was isolated from a separate cohort of 36 patients with early (stage F0-F1) or advanced (stage F3-F4) NASH-related fibrosis (n = 18/group) and analyzed by way of QRT-PCR. In livers with advanced fibrosis, OPN gene expression was double that of livers with early fibrosis (Fig. 6C). Additional analysis was conducted using RNA and protein harvested from explanted livers with NASH-related cirrhosis and residual tissue from nondiseased donor livers (n = 6/disease). Livers with NASH-related cirrhosis contained over 10 times more OPN protein (Fig. 6D; Supporting Information Fig. 4) and 5 times more OPN mRNA compared with nondiseased control livers (Fig. 7A).